艾灸督脉通过调控自噬受体介导的线粒体自噬功能改善APP/PS1小鼠学习记忆能力OA

Objective To investigate the regulatory role and mechanism of moxibustion at governor vessel acupoints on mitoph-agy in APP/PS1 transgenic mice with Alzheimer's disease(AD).Methods Adaptive feeding for 1 week and screening based on the water maze test were performed for 62 male APP/PS1 transgenic mice aged 5 months,among which 2 were excluded,and the remaining 60 mice were randomly divided into model group,moxibustion group,moxibustion+inhibitor group,and moxi-bustion+agonist group,with 15 mice in each group;another 15 C57BL/6J mice of the same age were selected as control group.For the mice in the moxibustion group,the moxibustion+inhibitor group,and the moxibustion+agonist group,press-ing moxibustion with Radix Aconiti Lateralis Preparata cake was performed at Baihui point for 20 minutes,while suspended moxibustion was performed at Fengfu and Dazhui points for 10 minutes at each acupoint,once a day for 6 days a week,fol-lowed by 1 day of rest,for a treatment course of 4 consecutive weeks.The mice in the model group and the control group were fixed alone without any intervention.The Morris water maze test was used to measure escape latency;HE staining was used to observe the histomorphological changes of the hippocampus;transmission electron microscopy was used to observe the number and morphology of mitochondria and autophagosome in the hippocampal CA1 region;immunohistochemistry was used to meas-ure the mean absorbance value of the protein expression of β-amyloid(Aβ)1-42;the JC-1 mitochondrial membrane potential assay kit was used to measure the loss of mitochondrial membrane potential in cells in hippocampal tissue;Western blot was used to measure the protein expression levels of BCL2/adenovirus E1B 19 kDa-interacting protein 3(BNIP3),adenovirus E1B 19 kDa-interacting protein 3-like(BNIP3L),FUN14 domain-containing protein 1(FUNDC1),prohibitin 2(PHB2),microtubule-associ-ated protein 1 light chain 3B-Ⅰ(LC3B-Ⅰ),microtubule-associated protein 1 light chain 3B-Ⅱ(LC3B-Ⅱ),and mammalian tar-get of rapamycin(mTOR)in hippocampal tissue,and RT-qPCR was used to measure the mRNA expression levels of BNIP3,BNIP3L,FUNDC1,PHB2,and mTOR in hippocampal tissue.Results Compared with the control group,the model group had a significant increase in escape latency(P＜0.05),a reduction in the number of hippocampal neurons,disordered arrangement of hippocampal neurons,mitochondrial ultrastructure damage,and an increase in the number of autophagosomes,as well as a significant increase in the mean absorbance value of the positive expression of Aβ1-42 protein in hippocampal tissue(P＜0.05),a significant reduction in mitochondrial membrane potential(P＜0.05),significant reductions in the protein and mRNA expres-sion levels of BNIP3,BNIP3L,FUNDC1,and PHB2(P＜0.05),significant reductions in the protein expression levels of LC3B-Ⅰ/LC3B-Ⅱ(P＜0.05),and significant increases in the protein and mRNA expression levels of mTOR(P＜0.05).Compared with the model group,the moxibustion group and the moxibustion+agonist group had a significant reduction in es-cape latency(P＜0.05),improvements in the number and arrangement of hippocampal neurons and mitochondrial ultrastruc-ture,and a significant reduction in the number of autophagosomes,as well as a significant reduction in the mean absorbance value of the positive expression of Aβ1-42 protein in hippocampal tissue(P＜0.05),a significant increase in mitochondrial mem-brane potential(P＜0.05),significant increases in the protein and mRNA expression levels of BNIP3,BNIP3L,FUNDC1,and PHB2(P＜0.05),significant increases in the protein expression levels of LC3B-Ⅰ/LC3B-Ⅱ(P＜0.05),and significant re-ductions in the protein and mRNA expression levels of mTOR(P＜0.05).Conclusion Moxibustion at governor vessel acu-points can improve learning and memory abilities in APP/PS1 mice,possibly by inhibiting the expression of mTOR and promo-ting the expression of the mitophagy receptors BNIP3,BNIP3L,FUNDC1,and PHB2,thereby mediating mitophagy,maintai-ning mitochondrial homeostasis,and alleviating Aβ-related neuronal injury.